CN105158129B - A kind of erythrocyte sedimentation rate (ESR) standard items and preparation method thereof - Google Patents

Abstract

A standard product of erythrocyte sedimentation rate and a preparation method thereof, including imitation human erythrocytes, a protective agent (liquid A) and a preservative (liquid B) placed in a Widman's erythrocyte sedimentation tube, the preparation method is as follows; Step1, take the tested The fresh blood of domestic animals qualified for quarantine is anticoagulated with 6:1 EDTA, and the supernatant is removed after centrifugation for 5 minutes; Step2, the bottom cells after centrifugation are washed with solution A, and then centrifuged again, and the bottom red blood cells are retained; Step3, the Add 20 times of B solution to the reserved red blood cells and mix thoroughly for 1 hour, and remove the supernatant after standing for 24 hours; Step4, then mix the collected and prepared mixture in A solution at a ratio of 1:1 to make a mock Human red blood cell granules, through the flow cytometer or hematology analyzer, calculate the exact cell number; Step5, inject the human red blood cell granules and sodium citrate in different proportions and concentrations in a 1:1 ratio mixture At the scale inside the Weisser's ESR tube, let it stand at room temperature for 1 hour, and then seal it for storage.

Description

A kind of erythrocyte sedimentation rate standard substance and preparation method thereof

technical field

The invention relates to the medical field, in particular to an erythrocyte sedimentation rate standard product and a preparation method thereof.

Background technique

Erythrocyte sedimentation rate, referred to as erythrocyte sedimentation rate, refers to the natural sinking rate of red blood cells in anticoagulated whole blood in vitro under specified conditions. The erythrocyte sedimentation standard is the metering substance for the determination of the erythrocyte sedimentation analyzer, operating technology, and quantitative analysis of the traceability of the environment. It is the only basis for the evaluation of the quality control of the room and the outside of the erythrocyte sedimentation rate analysis and to ensure the accuracy and precision of clinical test results. , but also according to user needs for high, medium and low value. At present, only some advanced countries in the world have such products. The preparation cost is high and the standard product has only one value.

Contents of the invention

In order to solve the above technical problems, the object of the present invention is to provide a method to solve the existing defects.

In order to achieve the above object, one kind of erythrocyte sedimentation rate standard product of the present invention is characterized in that: described erythrocyte sedimentation rate standard product comprises imitation human erythrocyte, cell activity protective agent (A liquid) and cell Active preservative (B liquid); the composition ratio of described cell activity protection liquid (A liquid) is, glucose 15-25g/L, boric acid 2-6g/L, sodium borate 1-2g/L, adenine 3 -5g/L, sodium fluoride 1-2g/L; The composition ratio of described cell viability preservation solution (B liquid) is, disodium hydrogen phosphate 36g/L, sodium dihydrogen carbonate 31g/L, glutaraldehyde 25% 25mg/L, propylene glycol 3g/L, calf serum 0.9g/L.

The preparation method of this erythrocyte sedimentation rate standard product is:

Step1, take the fresh slaughtered livestock fresh blood that has passed the inspection and quarantine and anticoagulate by volume 6:1 EDTA, centrifuge for 5 minutes and then remove the supernatant;

Step2, wash the bottom cells after centrifugation with a cell activity protection agent (solution A), then centrifuge the bottom cells again, repeat three times, and keep the bottom red blood cells;

Step3, add 20 times the cell viability preservative (solution B) to the reserved lower red blood cells and mix well for 1 hour, and remove the supernatant after standing for 24 hours;

Step4, then mix the collected and prepared red blood cells and the lower layer mixture of cell viability preserving agent (liquid B) in the ratio of 1:1 in the cell viability protecting agent (liquid A) to make human red blood cell granules, and pass flow cytometry Analyzer or hematology analyzer to calculate the exact number of cells;

Step5, the accurately counted human red blood cell granules and sodium citrate produced in Step4 were injected into the scale of the Weisserman's erythrocyte sedimentation tube according to different proportions, left at room temperature for 1 hour, and then sealed and stored.

Compared with the prior art, the present invention has the beneficial effects that: a kind of erythrocyte sedimentation rate standard product proposed by the present invention utilizes red blood cells in poultry blood to prepare imitation human red blood cells, and utilizes cell activity protective agent (A liquid), preservative (Liquid B), which simulates the osmotic pressure in human blood, and performs long-term maintenance and preservation of human red blood cells. The invention adopts the poultry blood that has been inspected and quarantined, has an economical source, is green and environmentally friendly, and has no biological safety hazards, completely replaces human red blood cells, has controllable stability, and can be prepared into standard products of erythrocyte sedimentation rate with different values according to clinical needs.

Description of drawings

Fig. 1 is the schematic diagram that the erythrocyte sedimentation rate standard product of the present invention is located in the Wei's method erythrocyte sedimentation tube;

The serial numbers and titles of the attached drawings: 1. liquid level, 2. erythrocyte layer.

Detailed ways

In order to describe the technical content, structural features, goals and effects of the present invention in detail, the following examples are given to describe the embodiments in conjunction with the accompanying drawings.

Example 1

Please refer to shown in Fig. 1, the present invention provides a kind of erythrocyte sedimentation rate standard product and preparation method thereof, at first utilize glucose 15g/L, boric acid 6g/L, sodium borate 2g/L, adenine 3g/L, sodium fluoride 1g /L is configured into a cell activity protection solution (liquid A), and then use disodium hydrogen phosphate 36g/L, sodium bicarbonate 31g/L, glutaraldehyde 25% 25mg/L, propylene glycol 3g/L, calf serum 0.9g /L to prepare cell activity preservation solution (B solution); then take the fresh blood of slaughtered livestock that has passed the inspection and quarantine and anticoagulate with 6:1 EDTA, centrifuge for 5 minutes, remove the supernatant, and wash the bottom cells with a protective agent , centrifugal precipitation, repeated three times, retain the red blood cells in the lower layer, add 20 times the preservative and mix well for 1 hour, after standing for 24 hours, remove the supernatant, collect the prepared mixture and mix it in the protective agent in a ratio of 1:1 ( Prepare humanoid red blood cells in liquid A); finally, inject the simulated human red blood cell particles accurately counted in the 1:1 ratio mixture into the scale of the Weisserman's ESR tube at the following concentration, let it stand at room temperature for 1 hour, and read the ESR tube The height of the descending layer of erythrocytes and the upper layer of transparent liquid is injected into 1, 2, 3, 4, and 5 erythrocyte sedimentation tubes according to the ratio in the table below and sealed up and down to obtain 5 kinds of standard products.

RBC granule mixture sodium citrate 0.2ml 1.8ml 0.4ml 1.6ml 0.6ml 1.4ml 0.8ml 1.2ml 1.0ml 1.0ml

Example 2

First, use glucose 25g/L, boric acid 5g/L, sodium borate 1g/L, adenine 5g/L, and sodium fluoride 2g/L to prepare a cell activity protection solution (A solution), then use disodium hydrogen phosphate 36g/L, Sodium bicarbonate 31g/L, glutaraldehyde 25% 25mg/L, propylene glycol 3g/L, and calf serum 0.9g/L were prepared into a cell viability preservation solution (B liquid); Anticoagulate the fresh blood of domestic animals with 6:1 EDTA, centrifuge for 5 minutes, remove the supernatant, wash the bottom cells with a protective agent, and centrifuge for three times, keep the lower red blood cells, add 20 times the preservative and mix well for 1 hour. After standing for 24 hours, remove the supernatant, collect the prepared mixture and mix it in the protective agent (solution A) at a ratio of 1:1 to make humanoid red blood cells; Red blood cell granules are injected into the scale of Wei's erythrocyte sedimentation tube according to the following concentration ratio, and stand at room temperature for 1 hour, read the height of the descending layer of erythrocytes in the erythrocyte sedimentation tube and the upper layer of transparent liquid, and inject 1, 2, 3, 4. Seal up and down the inside of the ESR tube to obtain 5 kinds of standard products.

RBC granule mixture sodium citrate 0.2ml 1.8ml 0.4ml 1.6ml 0.6ml 1.4ml 0.8ml 1.2ml 1.0ml 1.0ml

In summary, it is only a preferred embodiment of the present invention, and does not limit the protection scope of the present invention. All equivalent changes and modifications made according to the scope of the patent of the present invention and the content of the specification are all covered by the patent of the present invention. within.

Claims (2)

1. A standard product of erythrocyte sedimentation rate, characterized in that: said standard product of erythrocyte sedimentation rate comprises imitation human erythrocytes, cell viability protecting agent (A liquid), cell viability preserving agent (B liquid) ) and sodium citrate; the composition proportioning of described cell activity protection solution (A liquid) is, glucose 15-25g/L, boric acid 2-6g/L, sodium borate 1-2g/L, adenine 3- 5g/L, sodium fluoride 1-2g/L; the composition ratio of described cell viability preservation solution (B liquid) is, disodium hydrogen phosphate 36g/L, sodium dihydrogen carbonate 31g/L, glutaraldehyde 25g/L % 25mg/L, propylene glycol 3g/L, calf serum 0.9g/L. 2. a method for preparing erythrocyte sedimentation rate standard substance as claimed in claim 1, is characterized in that: Step1, take the fresh slaughtered livestock fresh blood that has passed the inspection and quarantine and anticoagulate by volume 6:1 EDTA, centrifuge for 5 minutes and then remove the supernatant; Step2, wash the bottom cells after centrifugation with a cell activity protection agent (solution A), then centrifuge the bottom cells again, repeat three times, and keep the bottom red blood cells; Step3, add 20 times the cell viability preservative (solution B) to the reserved lower red blood cells and mix well for 1 hour, and remove the supernatant after standing for 24 hours; Step4, then mix the lower layer mixture of the collected red blood cells and the cell viability preservation agent (B solution) in the ratio of 1:1 in the cell viability protectant (A solution) to make human red blood cell granules, and pass the flow cytometry Analyzer or hematology analyzer to calculate the exact number of cells; Step5, the accurately counted human red blood cell granules and sodium citrate produced in Step4 were injected into the scale of the Weisserman's erythrocyte sedimentation tube according to different proportions, left at room temperature for 1 hour, and then sealed and stored.